Functional delivery of a membrane protein into oocyte membranes using bicelles

Biochemistry. 2010 Feb 2;49(4):653-5. doi: 10.1021/bi902155t.

Abstract

Voltage-gated potassium channel modulatory membrane protein KCNE3 was overexpressed and purified into both micelles and bicelles. Remarkably, microinjection of KCNE3 in bicelles into Xenopus oocytes resulted in functional co-assembly with the human KCNQ1 channel expressed therein. Microinjection of LMPC micelles containing KCNE3 did not result in channel modulation, indicating that bicelles sometimes succeed at delivering a membrane protein into a cellular membrane when classical micelles fail. Backbone NMR resonance assignments were completed for KCNE3 in both bicelles and LMPC, indicating that the secondary structure distribution in KCNE3's N-terminus and transmembrane domains exhibits only modest differences from that of KCNE1, even though these KCNE family members have very different effects on KCNQ1 channel function.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Humans
  • KCNQ1 Potassium Channel / chemistry
  • KCNQ1 Potassium Channel / metabolism
  • Membrane Potentials
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism*
  • Micelles
  • Microinjections
  • Models, Biological
  • Molecular Sequence Data
  • Oocytes / metabolism*
  • Potassium Channels, Voltage-Gated / chemistry
  • Potassium Channels, Voltage-Gated / metabolism*
  • Xenopus / metabolism

Substances

  • KCNE3 protein, human
  • KCNQ1 Potassium Channel
  • KCNQ1 protein, human
  • Membrane Proteins
  • Micelles
  • Potassium Channels, Voltage-Gated